ABSTRACT Improving Anticoagulation Monitoring in Pediatric Patients: Use of a Microfluidic Platform to Test Low Volume Blood Samples Obtained by Heel-Stick Collection (Fast-Track SBIR) The goal of this Fast-Track SBIR project is to develop a near patient platform for laboratory monitoring of anticoagulants (heparin, heparinoids, and oral direct Factor Xa inhibitors) in children utilizing blood obtained from heel-stick or finger-prick. Currently, over 2 mL of whole blood is required to perform the standard laboratory assays during anticoagulation monitoring, and sample collection typically necessitates invasive venipuncture. Due to the increase in blood sampling to monitor ongoing therapy for children and infants being treated with anticoagulants, there is a critical unmet need for minimally-invasive blood sampling methods and miniaturized blood tests that require small sample volumes. Baebies Inc. proposes to collaborate with Dr. Sitaram Emani from Boston Children's Hospital to develop a microfluidic platform for near patient laboratory monitoring of anticoagulation using blood collected by minimally invasive transcutaneous puncture. This technology will be novel and innovative in that it is the first device to perform anticoagulation monitoring on an ultra-small volume of blood collected by heel-stick or finger-prick. The product will simultaneously perform three key assays: anti-thrombin function (ATIII), Factor Xa activity (FXa) and FXa supplemented with exogenous ATIII. The small volume format inherent to digital microfluidics enables several biochemical assays to be performed on 50 L or less of whole blood and is perfectly suited for use in pediatric settings where frequent blood draws significantly increase the risk of iatrogenic anemia. By combining three technically complex assays into a single, automated platform, we can reduce sample-to- answer time and personnel time needed to perform each individual assay. The aims of Phase I are to: 1) translate FXa and ATIII assays onto the FINDER? digital microfluidic platform; 2) multiplex 3 assays on a single cartridge; and 3) perform initial analytical validation. The key milestone and metric for progression to Phase II will be successful demonstration of multiplexed assays on one cartridge with high reliability and precision. The Aims for Phase II are to: 1) utilize discarded whole blood samples to perform analytical validation; 2) optimize techniques for blood sample collection to minimize contact activation; and 3) perform preliminary clinical validation of near patient, minimally-invasive, low-volume sample testing by comparing to standard clinical values obtained by venipuncture. The research will be performed on the Baebies' FINDER platform, a compact near patient platform currently under final development. At the conclusion of Phase II, we will have a commercializable diagnostic product for comprehensive management of anticoagulation therapies using low volume whole blood samples in a hospital or clinic setting. Our final product will be submitted for FDA approval and will initially be marketed for use in pediatric patients in U.S. hospitals and clinics with a future market towards other patients who may experience the disadvantages of current testing methodologies (serial venipuncture and iatrogenic anemia).